|
| Status |
Public on Nov 01, 2017 |
| Title |
CD8 Effector Memory T cells, donor 6 |
| Sample type |
RNA |
| |
|
| Source name |
CD27-CD45RA- CD8+ T cell
|
| Organism |
Homo sapiens |
| Characteristics |
gender: female
|
| Treatment protocol |
Each T cell subsets was stimulated for 2 hours with 0.5µg/ml plate coated anti-CD3 (OKT3) and 5ng/ml rhIL-2 before RNA isolation. A final condition took stimulated CD8+ EMRA T cells, CD27-CD45RA+ and incubated them with a p38 MAPK inhibitor BIRB796 at a final concentration of 500nM. 0.5x106 cells were stimulated in each case.
|
| Growth protocol |
PBMC were isolated using Ficoll plaque. CD8+ T cells were purified by positive selection. Positively selected CD8+ T cells were labeled with CD27 FITC (M-T271) and CD45RA APC (HI100) and sorted into 4 subsets using a FACSAria.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA isolation was achived using the ARCTURUS PicoPure Isolation Kit (ThermoFisher). The concentration of small quantities of RNA was determined using the Nanodrop. Linear amplification of 10 ng of total RNA was performed using the Ovation Biotin RNA amplification
|
| Label |
Biotin
|
| Label protocol |
The Nugen Biotin labeling kit was used
|
| |
|
| Hybridization protocol |
Fragmented, labelled cDNA was hybridized to Affymetrix U133 plus 2 arrays according to the manufacturer’s protocol at the Central Biotechnology Service, Cardiff University, UK.
|
| Scan protocol |
Agilent GeneChip 3000 scanner
|
| Description |
Gene expression data from EM CD8+ T cells
|
| Data processing |
Raw intensity data from each microarray were normalized to expression values using RMAExpress. Gene Cluster 3.0 software was used to center the genes and cluster them using Kendall’s tau coefficient. Heat maps of the clustered data were created using Java TreeView software. Principle component analysis was generated using the Partek software package.
|
| |
|
| Submission date |
May 08, 2017 |
| Last update date |
Jan 23, 2018 |
| Contact name |
Sian M Henson |
| E-mail(s) |
s.henson@qmul.ac.uk
|
| Organization name |
QMUL
|
| Street address |
Charterhouse Square
|
| City |
London |
| ZIP/Postal code |
EC1M 6BQ |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE98640 |
Expression data from human CD8+ T cell subsets, defined using CD27 and CD45RA |
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